Conformation Selective Antibody Enables Genome Profiling and Leads to Discovery of Parallel G-Quadruplex in Human Telomeres

Cell Chem Biol. 2016 Oct 20;23(10):1261-1270. doi: 10.1016/j.chembiol.2016.08.013. Epub 2016 Sep 29.

Abstract

G-quadruplexes are specialized secondary structures in nucleic acids that possess significant conformational polymorphisms. The precise G-quadruplex conformations in vivo and their relevance to biological functions remain controversial and unclear, especially for telomeric G-quadruplexes. Here, we report a novel single-chain variable fragment (scFv) antibody, D1, with high binding selectivity for parallel G-quadruplexes in vitro and in vivo. Genome-wide chromatin immunoprecipitation using D1 and deep-sequencing revealed the consensus sequence for parallel G-quadruplex formation, which is characterized by G-rich sequence with a short loop size (<3 nt). By using D1, telomeric parallel G-quadruplex was identified and its formation was regulated by small molecular ligands targeting and telomere replication. Together, parallel G-quadruplex specific antibody D1 was found to be a valuable tool for determination of G-quadruplex and its conformation, which will prompt further studies on the structure of G-quadruplex and its biological implication in vivo.

Keywords: antibody; conformational selectivity; consensus sequence; parallel G-quadruplex; telomeric G-quadruplex.

MeSH terms

  • Base Sequence
  • Cell Line
  • Consensus Sequence
  • G-Quadruplexes*
  • Genome, Human
  • HeLa Cells
  • Humans
  • Ligands
  • Models, Molecular
  • Single-Chain Antibodies / chemistry*
  • Single-Chain Antibodies / immunology
  • Telomere / chemistry*
  • Telomere / genetics
  • Telomere / immunology

Substances

  • Ligands
  • Single-Chain Antibodies